DK161078B - ANALOGY PROCEDURE FOR THE PREPARATION OF THERAPEUTICALLY ACTIVE MITOSAN COMPOUNDS - Google Patents

Description

DK 161078 B

The present invention relates to an analogous process for the preparation of therapeutically active mitosan compounds having the general formula II defined in the preamble of claim 1, which is characterized by the characterizing part of claim 1. The present compounds are useful in the treatment of neoplastic disease states in animals.

In U.S. Patent Nos. 4,268,676 and Nos.

4,460,599 have been reported for research to find novel and useful compounds structurally related to the mitomycin, which have antibiotic activity, low toxicity and exhibit a significant degree of antitumor activity in animals.

More specifically, U.S. Patent No. 4,268,676 discloses compounds of the general formula O

Wherein Y is hydrogen or lower alkyl and X is a thiazolamino group, a furfurylamino group or a group of the formula: R 1 R - NC-R 3 R 2 30 1 2 where R, R and R are the same or various and is selected 3 from hydrogen and lower alkyl and R is selected from lower alkenyl, halo (lower) alkenyl, lower alkynyl, lower alkoxycarbonyl, thienyl, formamyl, tetrahydrofuryl 35 and benzenesulfonamide.

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DK 161078 B

Said U.S. patent also discloses treatment of neoplastic disease states in animals using a therapeutically effective amount of a compound of general formula Ia 5

ISLAND

II

CHO0CNHo O 2 2 0CHs 10

H3C 0 [_I ^ NY

wherein Y is hydrogen or lower alkyl and Z is a thiazolamino group, a furfurylamino group, a cyclopropyl-amino group, a pyridylamino group or a group of the formula:

R R

I I 7

- N-C - R

Wherein R, R and R are the same or different and are selected from among hydrogen and lower alkyl, and R is selected from lower alkenyl, halogeno (lower) alkenyl, lower alkynyl, lower alkoxycarbonyl, halogeno (lower) alkyl, hydroxy (lower) alkyl, pyridyl, thienyl, formamyl, tetrahydrofuryl, benzyl and benzenesulfonamide.

In U.S. Patent No. 2,837,383, a compound 6-N- (p-hydroxyphenyl) -mithomycin-C of the formula 30 no- ^ y -mA_ ^ η, ο-conh, XOr ·

CH> IIH 'NH

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DK 161078 B

3 having similar properties to the compounds mentioned above.

New mitosan compounds have now been provided which are advantageous compared to the known mitosan derivatives by showing a more favorable combination of antitumor activity and low toxicity.

Thus, the invention provides an analogous method for the preparation of therapeutically active mitosan compounds of the general formula

10 O

n 11 0 CH ~ OCNHo - 15 3 0 I Dny where Y is hydrogen or lower alkyl and X is a (lower) alkoxy-substituted quinolinylaminogroup · 20 or a (lower) alkyl-substituted thiazolamino group or a nitrogen-containing heterocyclic group selected from 1-piperazinyl-1,1-pyrrolinyl, 1-indolinyl, N-thiazoladinyl, N-morpholinyl and N-thiomorpholinyl groups, or a cyano, phenyl, carboxamido or (lower) alkoxycarbonyl substituted 1-aziridinyl group or a (lower) alkyl or formyl-substituted 1-piper azinyl group or a hydroxy-substituted piperidyl group or a (lower) alkoxy-substituted pyridylamino group or a carboxamido, mercapto or methylenedioxy substituted or

H

in a group of formula -N-R 'wherein R "is a nitrogen-containing heterocyclic group selected from quinuclidinyl, pyrazolyl, 1-triazolyl,

DK 161078B

4 isoquinolinyl, indazolyl, benzoxazolyl, thiadiazolyl and benzothiadiazolyl, and (lower) alkyl and halo-substituted derivatives thereof, or a butyrolactonyl group or an adamantyl group or a substituted lower alkyl group selected from mercapto (lower) alkyl, di- and tri- (lower) alkoxy- (lower) alkyl, (lower) alkylthio- (lower) alkyl and (lower) alkoxycarbonyl-substituted derivatives thereof, cyano- (lower) alkyl, mono-, di- and tri- (lower) alkoxyphenyl) - (lower) alkyl, phenylcyclo (lower) alkyl), 1-pyrrolidinyl (lower) alkyl, N- (lower) alkylpyrrolidinyl (lower) alkyl, and N-morpholinyl ( lower) alkyl.

Unless otherwise stated, the term "lower" used for "alkyl" groups must indicate straight or branched groups having 1-6 C atoms. For example, "lower alkyl" should mean and include methyl, ethyl, propyl, butyl, pentyl and hexyl groups as well as isopropyl groups, t-butyl groups and the like. Similarly, "lower" used for "alkoxy" must denote a group of 1-6 C atoms.

The analogous process of the invention is characterized in that the mytomycin A or N-alkyl-mitomycin A is reacted with a suitable amine and that the compound obtained is isolated from the reaction mixture.

The reactions used usually yield the desired product in the form of a crystalline solid which is readily dissolved in alcohol.

N-Alkylmitomycin A is prepared from mitomycin C, e.g. by the methods generally disclosed by Cheng et al., J. Med. Chem., 20, No. 6, 767-770 (1977).

The compounds of the present invention have anti-bacterial activity against Gram-positive and Gram-negative microorganisms in a manner similar to that found for the naturally occurring mitotic agents.

DK 161078 B

5 mycines and are thus potentially useful as therapeutic agents in the treatment of bacterial infections in humans and animals.

The utility of compounds of formula II 5 in antineoplastic therapeutic methods is evidenced by the results of in vivo test methods, where the compounds are used in varying dosage amounts on mice that have been induced with a P338 leukemia condition. The experiments were conducted according to "Lymphocytic Leukemia P338 - Protocol 10 1,200", published in Cancer Chemotherapy Reports, Part 3, Volume 3, no. 2, page 9 (September 1972). Briefly, the experiments involved the use of the test compound on 1 µF CDF female mice previously infected with 10 ascites cells implanted intraperitoneally. The test compounds were administered only on the first test day, and the animals were monitored, inter alia, for vitality, over a period of 35 days.

Test results for the compounds of Examples 1-37 are set forth in Table I. The data provided include optimal dose ("O.D."), i.e. the dose in mg / kg body weight of the animal at which the maximum therapeutic effects are consistently observed. Also included is the mean median survival time ("MST") expressed as MST for the test animals compared to MST for control animals x 100 ("% T / C" where T = Test and C = Control). . In the above-mentioned in vivo P338 test method, a% T / C value of 125 or higher indicates significant antineoplastic therapeutic activity. The lowest dose in mg / kg of body weight at which the value of 125 for the% T / C value is obtained is known as the least effective dose ("MED"). These doses are also listed in Table I. It should be noted that the exceptionally high MST values obtained by P338 tests reported in Table I are also indicative of the absence of substantial toxicity of the compounds at the indicated doses.

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Table I

Example Optimal dose of MST MED

mg / kg as% T / C

1 12.8 339 0.2 5 2 3.2 211 0.4 3 12.8 150 0.8 4 6.4 211 0.2 5 6.4 178 0.4 6 25.6 144 12.8 10 7 6.4 175 0.8 8 25.6 255 1.6 9 25.6 239 1.6 10 12.8 217 0.8 11 6.4 131 3.2 15 12 12.8 217 1.6 13 25.6 178 1.6 14 12.8 222 0.8 15 6.4 200 0.8 16 12.8 313 <0.2 20 17 6.4 172 0.4 18 6.4 134 1.6 19 3.2 167 <0.2 20 12.8 194 0.4 21 12.8 183 0.2 25 22 25.6 206 0.2 23 12.8 161 0.8 24 6.4 232 0.4 25 12.8 216 0.2 3Q 26 25.6 222 0.2 27 3.2 261 <0.2 28 25.6> 333 0.8 29 25.6 150 6.4 30 12.8 205 1.6 35 31 25.6 170 1.6 (continued)

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Table I (continued)

Example Optimal dose of MST MED

mg / kg as% T / C

32 12.8 205 0.8 5 33 25.6 132 6.4 34 12.8 172 3.2 35 25.6 188 1.6 36 25.6 200 6.4 37 12.8> 211 <0, 2 10

Among the most preferred of the compounds of the invention used as antineoplastic agents are those exhibiting a relative life-prolonging ability of more than twice the relative life-prolonging ability, which is generally characterized as evidence of clear therapeutic potential. , ie those with an MST value% T / C higher than 2 x 125. The class of such compounds is seen to comprise the compounds of Examples 1, 8, 16, 27 and 28.

As shown in Table I, initial single doses as small as 0.2 mg / kg showed significant antineoplastic long-term efficacy. Accordingly, in therapeutic treatment, unit doses as small as 0.001 mg or as large as 5 mg, preferably from 0.004 mg to 1.0 mg, of the compounds can be used as active ingredient in a suitable pharmaceutical composition. Such preparations can be used in a daily program requiring from 0.1 mg to 100 mg per day. preferably from 0.2 to 51.2 mg per kg. kg body weight of the animal suffering from neoplastic disease. It is preferred that the compounds are used parenterally. Suitable pharmaceutical compositions may comprise simple aqueous solutions of one or more of the compounds of formula II, but may also include well known pharmaceutically acceptable diluents, adjuvants and / or carriers, such as medical salt.

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DK 1610 78 B

The method according to the invention is described in more detail in the following examples. Unless otherwise indicated, all reactions are carried out at room temperature (20 ° C) without further heating. Unless otherwise indicated, all thin-layer chromatographic methods (TLC) used to monitor the reaction procedures included the use of a pre-coated silica gel plate and a mixture of methanol and chloroform (2: 8 by volume) as the developing solvent.

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Example 1 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2-cyano-1-aziridinyl) -azirino [2 4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

A solution of mitomycin A (100 mg or 0.286 mmol) in 8 ml of anhydrous methanol was treated with 2-cyano-aziridine (38.9 mg or 0.572 mmol) and 30 mg of potassium carbonate under nitrogen at room temperature. When thin layer chromatograph on silica gel (2: 8 methanol-chloroform as solvent) showed that starting material was no longer available, the mixture was diluted with 50 ml of methylene chloride, filtered and evaporated under reduced pressure. The residue was purified by preparative thin layer chromatography on silica gel with a mixture of methanol 25 and chloroform (2: 8 by volume) as solvent.

This process yielded 33 mg (30% yield) of the desired product, m.p. 87-89 ° C (dec.) And with the following analysis: NMR (CDCl 3, TS): δ values in ppm.

The disappearance of a singlet at 4.02 (derived from the 6-methoxy group in the starting material) and the emergence of new signals at 2.13 (d, 2) and 2.53 (wide s, 1).

Example 2 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-35-methyl-6- (thiomorpholinyl) -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

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9

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 52 mg mitomycin A and 500 mg thiomorpholine, 14 mg (22% yield) of the desired product was obtained with m.p. 90-91 ° C (dec.) And the following analysis: NMR (CDCl3 + TS) δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and gain peak at 2.8 (m, 4 times increase) and 3.6 (m, 4 times increase).

Example 3 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (1-indolinyl) -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] -15 indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 69 mg of indoline 45 mg (36% yield) of the desired product was obtained with m.p. 127-135 ° C (dec.) And the following analysis: NMR (CDCl3, TS) δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 2.85-3.7 (group, 4) and 6.15-25 7f5 (group, 4).

Example 4 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6 - [(6-methoxy-3-pyridyl) amino] -azirino [21,31 : 3,4] -30 pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 2 drops of 3-amino-6-methoxy-pyridine, 96 mg (76% yield) of the desired product was obtained, m.p. 260-262 ° C (dec.). and the following analysis;

DK 161078 B

NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 3.93 (s, 3), 6.77 (s, 1), 7.26 (d, 1), 7.60 (d, 1) and 7.87 (s, 1).

5

Example 5 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6 - [(6-methoxy-8-quinolinyl) amino] -azirino [2 ', 3 ': 3,4] pyrrolo [1,2-a] indole carbamate.

This compound was prepared by the procedure described in Example 1. From 60 mg of mitomycin A and 54 mg of 8-amino-6-methoxyquinoline, 26 mg (32% yield) of the desired product was obtained, m.p. 135-145 ° C (dec) and the following analysis: 15 NMR (CDCl3, TS) δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of new spikes at 6.4 (d, 1), 6.67 (d, 1), 7.30 (dd, 1), 8.0 (dd, 1) and 8.90 (dd, 1).

Example 6 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (3-quinuclidinylamino) -azirino [2 ', 3': 3.4 ] pyrrol-lo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 3-aminoquinuclidine (prepared by treating an aqueous solution of 73 mg of 3-aminoquinuclidine hydrochloride with sodium hydroxide), 86 mg (54% yield) of the desired product was obtained, m.p. 138-146 ° C (dec.) And the following analysis: NMR (CDCl3, TS) δ values in ppm.

Absence of the 6-methoxy tip at 4.02, amplification of the peaks at 2.8 and 3.8 and the emergence of new 35 wide peaks at 1.2 and 2.5.

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Example 7 1,13,2,8,83,8b-Hex3hydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [2- (γ-butyrolactonyl) amino] azirino [2 ', 31: 3,4] -pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 60 mg of α-amino-γ-butyrolactone hydrochloride, 68 mg (57% yield) of the desired product was obtained, m.p. 87-89 ° C (dec.) And the following analysis: NMR (DMSO-dg, TS) δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 1.90-2.87 (m, 2), 3.80-4.70 (m, 3) and 8.3-9.2 (wide s, 1).

Example 8 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (4-carboxamidoanilino) -azirino [21,31: 3,4] pyrro -LO [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 100 mg of mitomycin A and 82 mg of 4-aminobenzamide, 36 mg (28% yield) of the desired product was obtained with m.p. 167-169 ° C (dec.) And the following analysis: NMR (Acetone-dg, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of new points at 6.67 (d, 3) and 7.73 (d, 2).

Example 9 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-30methyl-6 "(3,4-dimethoxybenzylamino) -azirino [2,3,3 ': 3 , 4] pyrrolo [^ ,. 2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 29 mg of mitomycin A and 69.4 mg of 3,4-dimethoxybenzylamine, 29 mg (72% yield) of the desired product was obtained with m.p. 112 ° C (dec) and the following analysis:

DK 161078 B

12 NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 3.9 (s, 6), 4.65-4.75 (d, 2), 6.55 (wide s, 1) and 6 , 86 (s, 3).

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Example 10 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6 - [(1-ethyl-2-pyrrolidino) methylamino] -azirino [2 ', 31: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 150 mg of mitomycin A and 2 drops of 2-aminomethyl-1-ethylpyrrolidine, 78 mg (41% yield) of the desired product decomposed at temperatures above 300 ° C were obtained and showed the following analysis: NMR (CDCl ^ r TS): 6 values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new spikes at 1.07 (t, 3), 1.4-2.33 (m, 5), 2.36-3.03 (m, 4), 3.3-3.83 (m, 2) and 6.77-7.20 (wide s, 1).

Example 11 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [(1-methoxycarbonyl-3-methylthio) propylamino] -azirino [ 2 *, 3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was replaced with 0.5 ml of triethylamine.

From 150 mg of mitomycin A and 110 mg of L-methionine methyl ester hydrochloride, 64 mg (30% of yield) of the desired product was obtained, m.p. 83-85 ° C (dec.) And the following analysis: NMR (CDCl3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of new peaks at 1.63-2.40 (m, 3), 2.10 (s, 3), 2.43-3.0 (m, 2), 3.80 (s, 3) and 8.3-9.3 (wide s,]).

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Example 12 1,13,2,8,8a, 8b-Hexahydro-8- (hydroxymethyl). 8a-methoxy-5-methyl-6- (2-phenylcyclopropylamino) -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 125 mg of mitomycin A and 85 mg of 2-phenylcyclopropylamine, 70 mg (63%) of the desired product decomposed at temperatures 10 above 250 ° C were obtained and showed the following analysis: NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 0.6-1.53 (m, 4), 6.20-6.50 (wide s, 1) and 7.18 (wide s) , 5).

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Example 13 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6 - [(5-chloro-2-benzoxazolyl) amino] -azirino [2 ', 3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 100 mg of mitomycin A and 50 mg of 2-amino-5-chlorobenzoxazole, 35 mg (25% yield) of the desired product was obtained, m.p. 118-120 ° C (dec) and the following analysis: 25 NMR (CDCl-3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new peaks in the range of 6.70-7.63 (m, 4).

Example 14 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [5-methyl-2- (1,3,4-thiadiazolyl) amino ] -azirino- [2 ', 3': 3,4] pyrrolo (1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 100 mg of mitomycin A

33 and 53 mg of 2-amino-5-methyl-1,3,4-thiadiazole were obtained 31 mg (25% yield) of the desired product, m.p. 91-93 ° C (dec.) And the following analysis: 14

DK 161078 B

NMR (CDCl3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 2.68 (s, 3) and 7.47-7.63 (wide s, 1).

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Example 15 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2,2-dimethoxyethylamino) -azirino E2,3,3,4,4 ] - pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 60 mg of mitomycin A and 35 mg of 2,2-dimethoxyethylamine, 60 mg (83% yield) of the desired product was decomposed, decomposing at 15 temperatures above 220 ° C and showing the following analysis: NMR (CDCl3 + TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 3.45 (s, 6), 3.33-3.93 (m, 2), 4.33-4.85 (wide s, 1) and 6.15-6.66 (wide s, 1).

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Example 16 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2-mercaptoethylamino) -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that 0.5 ml of triethylamine was used instead of the potassium carbonate.

From 150 mg of mitomycin A and 100 mg of 2-mercaptoethylamine hydrochloride, 50 mg (44% yield) of the desired product was obtained with m.p. 152-154 ° C (dec) and the following analysis: NMR (DMSO-dg, TMS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 2.53-3.10 (m, 4), 7.30-7.50 (wide s, 1).

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Example 17 1,13,2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6 - [(4-methyl-2-thiazolyl) amino] -azirino [2 ', 31: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 150 mg of mitomycin A and 96 mg of 2-amino-4-methylthiazole, 85 mg (59% yield) of the desired product was obtained, m.p. 116-118 ° C (dec.) And the following analysis: 10 NMR (CDCl 3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new spikes at 2.23 (s, 3), 6.30-6.60 (wide s, 1), and 7.30 (s, 1).

Example 18 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (4-mercaptoanilino) -azirino [21,3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 200 mg of mitomycin A and 143 mg of 4-mercaptoaniline, 120 mg (47% yield) of the desired product was obtained with m.p. 97-99 ° C (dec.) And the following analysis: NMR (CDCl3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of new points at 6.53 (d, 2) and 7.0-7.7 (m, 3).

Example 19 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-30-methyl-6- (3,4-methylenedioxyanilino) -azirino [2 ', 3': 3 , 4] pyrrolo [l, 2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 80 mg of mitomycin A 35 and 0.1 ml of 3,4-methylenedioxyaniline, 50 mg (48% yield) of the desired product was obtained with m.p. 86-88 ° C (dec.) And the following analysis: 4 * 16

DK 16107 8 B

NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 5.97 (s, 2), 6.0-6.7 (m, 3), 7.27 (s, 1).

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Example 20 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [2- (1-pyrrolidino) ethylamino] -azirino [21,3 '; 3,4] pyrrolo [l, 2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 0.2 ml of 2- (1-pyrrolidino) -ethylamine, 7.5 mg (61% yield) of the desired product was decomposed, decomposing at 15 temperatures above 200 ° C and showing the following analysis: NMR (CDCl-., TS): δ values in ppm.

^ /

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 1.57-1.93 (m, 4), 2.33-3.03 (m, 8) and 6.92 (t, 1 ).

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Example 21 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (5-isoquinolinylamino) -azirino [2 ', 31: 3,4] pyrro -LO [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 90 mg of mitomycin A and 810 mg of 5-aminoisoquinoline, 28 mg (24% yield) of the desired product was obtained without a melting point below 340 ° C and with the following analysis: 30 NMR (GDClg, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new peaks at 6.8-7.65 (m, 3), 7.85 (d, 1) and 8.55 (d, 1).

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Example 22 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (5-indazolylamino) -azirino [2 *, 3 *: 3.4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 90 mg of mitomycin A and 666 mg of 5-aminoindazole 35 mg (30% yield) of the desired product was obtained without a melting point below 340 ° C and with the following analysis: 10 NMR (CDClg, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 6.8-7.65 (m, 3) and 8.0 (s, 1).

Example 23 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [4- (2,1,3-benzothiadiazolyl) amino] -azirino [ 2 ', 31; 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. The reaction did not proceed to the 2Q end within 19 hours despite the use of excess amine. From 50 mg of mitomycin A and 300 mg of 4-amino-2,1,3-benzothiadiazole, 32 mg (48% yield) of the desired product was obtained with m.p. 139-140 ° C (dec) and the following analysis: 25 NMR (CDClCl + CD ^ OD, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new peaks at 6.6 (m, 1), 7.6 (m, 2) and 8.25 (wide s, 1).

Example 24 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2-cyanoethylamino) -azirino [2 ', 3': 3.4 ] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1, with the exception that 0.5 ml of triethylamine was used instead of the potassium carbonate.

From 210 mg of mitomycin A and 90 mg of 3-aminopropionitrile fumarate 151 mg (65% yield) of the desired product was obtained with m.p. 68-70 ° C (dec.) And the following analysis: NMR (CDCl 3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new peaks at 2.1-2.77 (m, 4) and 6.57 (t, 1).

Example 2 5 1 q 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [1- (3-pyrrolinyl)] azirino [2 ', 3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted and a change was required by the presence of pyrrolidine impurity in the 3-pyrroline commercial product. The pyrrolidine formed a crystalline derivative with mitomycin A which was removed from the mixture by filtration. The filtrate was then worked up as described in Example 1. From 100 mg of mitomycin A and 1 g of 3-pyrroline commercial product, 30 mg (27% yield) of the desired product was obtained with a partial decomposition temperature of 85-90 ° C. but without melting point below 250 ° C, and which showed the following analysis: 25 NMR (CDCl3, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of a new peak at 5.9 (s, 2). It was not possible to distinguish the 2-proton peak in the 3.4 range from other absorption.

30

Example 26 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8-methoxy-5-methyl-6- (3-thiazolidino) -azorino [2 ', 3'; 3,4] pyrrole [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 250 mg mitomycin Ά

DK 161078 B

19 and 0.5 ml of thiazolidine were obtained 125 mg (43% yield) of the desired product, m.p. 105-107 ° C (dec) and the following analysis: NMR (CDCl3, TMS): δ values in ppm.

Absence of the 6-methoxy tip at 4.02 and the emergence of new tips at 2.62 (wide s, 2), 2.68-3.02 (wide s, 2) and 3.32-4.02 ( wide s, 2).

Example 27 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [1- (4-methylpiperazino)] - azirino [2 ', 3' : 3,4] pyr rolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 0.2 ml of N-methylpiperazine, 50 mg (42% yield) of the desired product was obtained with m.p. 84-87 ° C (dec.) And the following analysis: NMR (CDClgj TS TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of new peaks at 2.27 (s, 3), 2.47 (t, 4) and 2.92 (t, 4).

Example 28 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [3- (pyrazolyl) amino] azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 100 mg of mitomycin A and 48 mg of 3-aminopyrazole, 50 mg (44% yield) of the desired product was obtained with m.p. 142-145 ° C (dec) and the following analysis: NMR (CDCl 3, TMS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and the emergence of new spikes at 6.50 (d, 2), 6.67-6.83 (wide s, 1) and 8.07 (s, 1).

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20

Example 29 1,13,2,8,83,8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [2- (N-morpholino) ethylamino] -azino [21 , 31: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by process I

as described in Example 1 except that the potassium carbonate was omitted. Dd from 100 mg mitomycin A j and 0.5 ml of N- (2-aminoethyl) morpholine was obtained 70 mg (55% j yield) of the desired product, m.p. 74-76 ° C (dec.) And the following analysis: NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 2.27-2.73 (wide, 8), 3.47-4.03 (wide, 4) and 7.27 (t, 1 ).

15

Example 30 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [2- (ethylthio) ethylamino] -azino [2,3,3 , 45-pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1, except that 0.5 ml of triethylamine was used instead of the potassium carbonate.

From 250 mg of mitomycin A and 101.5 mg of 2- (ethylthio) ethylamine hydrochloride, 220 mg (73% yield) of the desired product was obtained, m.p. 103-106 ° C (dec.) And the following analysis: NMR (CDCl 3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 1.27 (t, 3), 2.40-2.90 (m, 4), 3.40-3.93 (m, 2) and 6.56 (t, 1).

Example 31 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-1,5-dimethyl-6- (2-mercaptoethylamino) -azirino [2 ', 3': 3, 4] -35 pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 with the exception that 0.5

DK 161078 B

21 ml of triethylamine was used instead of the potassium carbonate.

From 250 mg of N-methyl-mitomycin A and 78 mg of 2-mercaptoethylamine hydrochloride, 150 mg (54% yield) of the desired product was obtained with m.p. 85-87 ° C (dec.) And following analysis: NMR (CDCl3, TS): δ values in ppm.

Absence of the 6-methoxy group at 4.05 and emergence of new spikes at 2.57-3.10 (wide s, 4) and 6.20- 6.93 (wide s, 1).

10

Example 32 1,1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2-methoxyethylamino) azirino in 2 ', 3': 3,4] pyrrolo - [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 120 mg of mitomycin A and 0.2 ml of 2-methoxyethylamine, 99 mg (73% yield) of the desired product was obtained with m.p. 106-109 ° C (dec) and the following analysis: NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 3.42 (s, 3), 3.5-3.9 (wide s, 4) and 6.27-6.77 (wide s) , 1).

Example 33 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (1-adamantylamino) -azirino [21,31: 3,4] pyrrolo - [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. The reaction was not complete within 48 hours despite the use of excess amine. From 147 mg of mitomycin A and 666 mg of 1-aminoadamantane, 60 mg (30% yield) of the desired product was obtained with m.p. 149-150 ° C (dec.), Partial decomposition at 85-90 ° C, and the following analysis: NMR (CDCl3 + CD3OD, TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 1.55-2.3 (m, 15).

i *

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22

Example 34 1 / 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [1- (1,3,4-triazolyl) amino] -azirino [2 ', 3': 3,4] -pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1. From 100 mg of mitomycin A and 80 mg of 1-amino-1,3,4-triazole, 35 mg (30% yield) of the desired product was obtained with m.p. > 250 ° C (dec) and the following analysis; 1 C NMR (CDCl 3 / TMS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 8.00 (s, 2).

Example 35 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (3,4,5-trimethoxybenzylamino) -azirino [21,3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that the potassium carbonate was omitted. From 65 mg of mitomycin A and 437 mg of 3,4,5-trimethoxybenzylamine 55 mg (57% yield) of the desired product were obtained with m.p. 94-95 ° C (dec.) And the following analysis: NMR (CDCl3 / TS); δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 3.85 (s, 9), 4.46-4.76 (d, 2) and 6.45 (s, 2).

Example 36 3C 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-1,5-dimethyl-6- [2- (ethylthio) ethylamino] -azirino [2 ', 3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1 except that 0.5 35 ml of triethylamine was used instead of the potassium carbonate.

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23

From 120 mg of N-methyl-mitomycin A and 70 mg of 2- (ethylchio) -ethylamine hydrochloride, 100 mg (69% yield) of the desired product was obtained, m.p. 114-116 ° C (dec.) And the following analysis: R NMR (CDCl-cf TS): δ values in ppm.

Absence of the 6-methoxy peak at 4.02 and emergence of new tips at 1.27 (t, 3), 2.40-2.93 (m, 4), 3.40-3.93 (m, 2 ) and 6.50-6.80 (wide s, 1).

Example 37 1, 1a, 2,8,8a, 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [1- (3-hydroxypiperidyl)] - azirino [2 ', 3' : 3,4] pyr rolo [1,2-a] indole-4,7-dione carbamate.

This compound was prepared by the procedure described in Example 1, except that 0.5 ml of triethylamine was used instead of the potassium carbonate.

From 130 mg of mitomycin A and 70 mg of 3-hydroxypiperidine hydrochloride, 80 mg (58% yield) of the desired product was obtained with m.p. 98-101 ° C (dec) and the following analysis: 20 NMR (CDCl3, TS): δ values in ppm.

Absence of 6-methoxy peak at 4.02 and emergence of new peak at 0.97-2.13 (wide m, 4), 2.17-3.13 (wide m, 4), 3.3-4 , 33 (wide m, 1) and 4.67-5.73 (wide s, 1).

25

Example 38

Using mitomycin A and the appropriate amine starting materials, the methods of the previous examples could be used in the preparation of the following compounds:

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(a) 1,1a, 2,8,8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-6- (2-phenyl-1-aziridinyl) -azirino [2 ', 3 *: 3.4 ] - pyrrolo [1,2-a] indole-4,7-dione carbamate, yield 37%, m.p. 160-167 ° C (dec), new NMR peaks: 1.16-2.1 δ (dd, 1), 2.2-2.9 (m, 2), 6.9-7.4 ( m, 5), (b) 1,1a, 2,8,8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-6- (2-methoxycarbonyl-1-aziridinyl) -azirino [2 ', 31 : 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate ,. yield 73%, m.p. 125-130 ° C (dec), new NMR peaks: 2.4-2.7 (m, 3), 3.73 (s, 3), (c) 1, 1a, 2.8.8a , 8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-6- (2-carboxamido-1-aziridinyl) -azirino [2 ', 3 1: 3,4] -pyrrolo [1,2-a] indole 4,7-dione carbamate, yield 47%, m.p. 80 ° C (dec), new NMR peaks: 2.25-2.75 (m, 3), 6.25-7.15 (d, 2), (d) 1, 1a, 2.8 , 8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-6- (N-morpholinyl) -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7 -dione carbamate, yield 95%, m.p. 200 ° C (dec.), New NMR peaks: 2.90 (t, 4), 3.74 (t, 4), (e) 1, 1a, 2,8,8a, 8b-hexahydro-8 (hydroxymethyl) -8- methoxy-6- (1-piperazinyl) -azirino [2 ', 31: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, yield 34% , m.p. 200 ° (dec.), New NMR peaks: 1.83 (wide s, 1), 2.96 (s, 8), (f) 1, 1a, 2,8,8a, 8b-hexahydro-8 (hydroxymethyl) -8a-methoxy-6- (4-formyl-1-piperazinyl) -azirino [2 ', 3': 3,4] -25 pyrrolo [1,2-a] indole-4,7-dione carbamate, yield 41%, m.p. 94-96 ° C (dec), new NMR peaks: 2.38-3.05 (m, 4), 3.06-3.88 (m, 4), 8.13 (s, 1).

Referring to the compounds represented by formula II, the above examples illustrate the following structural variations: 1. In the compounds of Examples 31 and 36, Y is lower alkyl and more particularly methyl. In all other examples, Y is hydrogen. The identity of Y is independent of the identity of X, cf. Examples 16 and 31, where 35 X is the same and Y is hydrogen and lower alkyl respectively.

See also Examples 30 and 36, which differ in the same way.

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2. Compounds in which X is a (lower) alkoxy-substituted quinolinylamino group or a (lower) alkyl-substituted thiazolylamino group are shown in Example 5 and Example 17, respectively.

3. Compounds in which X is a nitrogen-containing heterocyclic group selected from 1-pyrrolinyl, 1-indolinyl, N-thiazoladinyl, N-morpholinyl, 1-piperazinyl and N-thiomorpholinyl groups are shown, respectively. in Examples 25, 3, 26, 38 (d), 38 (e) and 2.

4. Compounds in which X is a cyano-, phenyl-, carboxamido- or (lower) alkoxycarbonyl-substituted 1-aziridinyl group are shown in Examples 1, 38 (a), 38 (c) and 38 (b), respectively.

5. Compounds in which X is a (lower) alkyl or formyl-substituted 1-piperazinyl group are shown in Examples 27 and 38 (f), respectively.

6. Compounds in which X is a hydroxy-substituted piperidyl group are shown in Example 37.

7. Compounds in which X is a (lower) alkoxy-substituted pyridylamino group are shown in Example 4.

8. Compounds in which X is a carboxamido, mer-capto or methylenedioxy-substituted anilino group are shown in Examples 8, 18 and 19, respectively.

Compounds wherein X is a group of the formula 25 and (lower) alkyl- and halogen-substituted derivatives thereof are shown, respectively, in Examples 6, 28, 34, 21, 22, 13, 14; and 23.

Compounds in which X is a group of the formula

H

f -N-R 'wherein R' is a butyrolactonyl group or an adamantyl group are shown in Examples 7 and 33, respectively.

Claims (5)

Compounds wherein X is a group of formula H -NR r, wherein R 'is a substituted lower alkyl group selected from mercapto- (lower) alkyl, carboxy- (lower) alkyl, mono-, di- and tri - (lower) alkoxy- (lower) alkyl, (lower) alkylthio- (lower) alkyl and (lower) alkoxycarbonyl-substituted derivatives thereof, cyano- (lower) alkyl, mono-, di- and tri- (lower) alkoxyphenyl (lower) alkyl, phenylcyclo (lower) alkyl, 1-pyrrolidinyl (lower) alkyl, N- (lower) alkylpyrrolidinyl (lower) alkyl and N-lowerpholinyl (lower) alkyl are shown in Examples 16, 31, 32, 15, 30, 36, 11, 24, 9, 35, 12, 20, 10 and 29. 15 1, Analogous Process for Preparing Therapeutically Active Mitosan Compounds of General Formula OX in? CH 2 OCNH 2 "wherein Y is hydrogen or lower alkyl and X is a (lower) alkoxy-substituted quinolinylamino group or (lower) alkyl-substituted thiazole amino group, or a nitrogen-containing heterocyclic group selected from 1-piperazinyl, 1 pyrrolinyl, 1-indolmyl, N-thiazoladinyl, N-morpholinyl and N-thiomorpholinyl groups, or a cyano, phenyl, carboxamido or (lower) alkoxycarbonyl-substituted 1-aziridinyl group or 35 lower) alkyl- or formyl-substituted 1-piperazinyl group or a hydroxy-substituted piperidyl group or a (lower) alkoxy-substituted pyridylamino group DK 161078 B or a carboxamido, mercapti or methylenedioxy-substituted anilino group or H -R 'wherein R "is a nitrogen-containing heterocyclic group selected from quinuclidinyl, pyrazolyl, 1-triazolyl, isoquinolinyl, indazolyl, benzoxazolyl, thiadiazolyl and benzothiadiazolyl, and 30 (lower) alkyl and halogen substituted derivatives or a butyrolactonyl group or an adamantyl group or a substituted lower alkyl group selected from the group consisting of mercapto (lower) alkyl, mono-, di- and tri (la) alkoxy- (lower) alkyl, (lower) alkylthio- (lower) ) alkyl and (lower) alkoxycarbonyl-substituted derivatives thereof, cyano- (lower) alkyl, mono-, di- and tri- (lower) alkoxyphenyl (lower) -20 alkyl, phenylcyclo (lower) alkyl, 1-pyrrolidinyl - (lower) alkyl, N- (lower) alkylpyrrolidinyl (lower) alkyl and N-morpholinyl (lower) alkyl * characterized by reacting mitomycin A or N-alkyl-mitomycin A with a suitable amine and 25 compound is isolated from the reaction mixture. A process according to claim 1 for the preparation of the compound 1,1a, 2,8,8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2-cyano-1-aziridinyl) ) -azino [21,3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that the compound is prepared from the appropriate starting materials. A process according to claim 1, for the preparation of the compound 1,1a, 2,8,8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (4-carboxamidoanilino) -azyric acid no [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that the compound is prepared from the appropriate starting materials. DK 161078 Β β ·> Process according to claim 1, for the preparation of the compound 1,1a, 2,8,8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (2-mercaptoethylamino) -azirone [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, 5th end, characterized in that the compound is prepared from the appropriate starting materials. Process according to claim 1, for the preparation of the compound 1,1a, 2,8,8a, 8b-hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [3- (pyrazolyl) amino] - azirino [2 ', 31: 3,4) pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that the compound is prepared from the appropriate starting materials.